Pathogenesis of reproductive failure induced by Trypanosoma vivax in experimentally infected pregnant ewes

The present study was aimed at investigating the effect of experimental infection by Trypanosoma vivax in different stages of pregnancy, determining the pathogenesis of reproductive failure, and confirming transplacental transmission. We used 12 pregnant ewes distributed into four experimental groups: G1, was formed by three ewes infected with T. vivax in the first third of pregnancy (30 days); G2 comprised three infected ewes in the final third of pregnancy (100 days); G3 and G4 were composed of three non-infected ewes with the same gestational period, respectively. Each ewe of G1 and G2 was inoculated with 1.25 × 10⁵ tripomastigotes. Clinical examination, determination of parasitemia, serum biochemistry (albumin, total protein, glucose, cholesterol, and urea), packed cell volume (PCV), serum progesterone, and pathological examination were performed. Placenta, amniotic fluid, blood and tissues from the fetuses and stillbirths were submitted to PCR. Two ewes of G1 (Ewe 1 and 3) presented severe infection and died in the 34^th and 35^th days post-infection (dpi), respectively; but both fetuses were recovered during necropsy. In G2, Ewe 5 aborted two fetuses on the 130^th day (30 dpi) of pregnancy; and Ewe 6 aborted one fetus in the 140^th day (40 dpi) of gestation. Ewes 2 and 4 delivered two weak lambs that died five days after birth. Factors possibly involved with the reproductive failure included high parasitemia, fever, low PCV, body score, serum glucose, total protein, cholesterol, and progesterone. Hepatitis, pericarditis, and encephalitis were observed in the aborted fetuses. The presence of T. vivax DNA in the placenta, amniotic fluid, blood, and tissues from the fetuses confirms the transplacental transmission of the parasite. Histological lesion in the fetuses and placenta also suggest the involvement of the parasite in the etiopathogenesis of reproductive failure in ewes.     

A homolog of the O157 urease-encoding O island 48 is present in porcine O149:H10 enterotoxigenic Escherichia coli

The relationship of the urease operon in the highly virulent O149 porcine enterotoxigenic Escherichia coli (ETEC) strain Ro8 to a genomic island (GI) homologous to O island (OI) 48 of O157 enterohemorrhagic E. coli (EHEC) strain EDL933 was investigated. Eighty-four of 84 O149:H10 strains were urease positive whereas 44 of 44 O149:H43 porcine ETEC strains were urease-negative. Seventeen of 17 O149:H10 strains that were tested possessed the OI-48 homolog whereas 24 of 24 O149:H43 strains lacked this OI. Transposon insertions in lipB or guaA genes in strain Ro8 eliminated urease activity while insertions in the caiF gene increased urease activity. When the O149 ure operon was cloned on a high copy number plasmid, urease expression was increased approximately 11-fold in Ro8 and 83-fold in O157 strain EDL933 compared with that in the wild type Ro8. The O149 urease activity was expressed despite the presence of the same premature stop codon in ureD that is present in ure+ O157:H7 strains that are urease-negative. The ure operon in Ro8 consists of 4 893 nucleotides with 99% identity with the ure operons in EHEC O157:H7 strains EDL933 and Sakai, and is part of a GI similar to GI-48 of strain EDL933. This OI, designated OI-48149 , is inserted in the serX tRNA gene in strain Ro8 and contains genes for urease, tellurite resistance, iha and an AIDA-I-like adhesin. The presence of a homolog of the O157:H7 OI-48 in highly virulent O149 porcine ETEC suggests that this OI may contribute to establishment of the bacteria in the intestine.     

Oxytocin Secretion and Management of Parturition in the Pig

This paper will examine the augmentation of oxytocin (OT) secretion in the final day before parturition in the pig, and review the key endocrine systems of influence. Recent experiments have shown that increases in plasma OT above baseline secretion were not seen until 7 hours prior to the birth of the first piglet. Progesterone release begins to decline some days before birth and is unlikely to provide such acute control of OT release immediately before birth. The involvement of relaxin is complex but it is possible that the prepartum relaxin surge, which ends close to the OT rise, attenuates OT secretion to prevent excessive myometrial activity until cervical softening has occurred. Increasing myometrial electromyographic activity appears to be seen as relaxin concentrations decline and OT increase. Endogenous opioid peptides within the hypothalamus exert a profound influence upon OT secretion during birth, and some effects are also seen prepartum. It has been proposed that this system may allow parturition to be interrupted or slowed in response to changes in environment. The therapeutic choice of exogenous injections of OT to control the onset time of parturition will be discussed in the light of current physiological knowledge.     

Virulence markers in Shiga toxin-producing Escherichia coli isolated from cattle.

This study identified potential virulence markers in 93 eae-positive and 179 eae-negative Shiga toxin-producing Escherichia coli (STEC), isolated from a random sampling of healthy cattle in southwestern Ontario. PCR amplification was used to identify genes for enterohemorrhagic E. coli (EHEC)-hemolysin, the EAF plasmid, and bundle-forming pili (Bfp); adherence to HEp-2 cells and to bovine colonocytes, and the fluorescent actin staining (FAS) test were used to characterize interaction of the bacteria with epithelial cells. The EHEC-hemolysin sequences were detected in 98% of eae-positive isolates compared with 34% of eae-negative isolates. All isolates were negative for EAF and bfp sequences. There was 100% correlation between localized adherence (LA) to HEp-2 cells and the FAS test. Forty-eight (52%) of the eae-positive isolates were LA/FAS-positive, whereas none of the 179 eae-negative isolates was positive in either test. Among the eae-negative isolates, 20 (11%) showed diffuse adherence and 5 (2.8%) showed enteroaggregative adherence to HEp-2 cells. Seventy-three percent of the eae-positive isolates adhered to bovine colonocytes, whereas only 26% of 120 eae-negative isolates that were tested adhered. All 13 O157:H7 isolates were positive for eae and EHEC-hemolysin gene sequences, LA/FAS, and adherence to bovine colonocytes. It is concluded that possession of genes for eae and EHEC hemolysin is correlated with the serotype of STEC, that production of EHEC hemolysin was highly correlated with serotypes implicated in human disease, and that none of the potential markers that were examined can be used to predict the potential virulence of an isolate.     

Impact of biodiversity-climate futures on primary production and metabolism in a model benthic estuarine system

Background  

Understanding the effects of anthropogenically-driven changes in global temperature, atmospheric carbon dioxide and biodiversity on the functionality of marine ecosystems is crucial for predicting and managing the associated impacts. Coastal ecosystems are important sources of carbon (primary production) to shelf waters and play a vital role in global nutrient cycling. These systems are especially vulnerable to the effects of human activities and will be the first areas impacted by rising sea levels. Within these coastal ecosystems, microalgal assemblages (microphytobenthos: MPB) are vital for autochthonous carbon fixation. The level of in situ production by MPB mediates the net carbon cycling of transitional ecosystems between net heterotrophic or autotrophic metabolism. In this study, we examine the interactive effects of elevated atmospheric CO₂ concentrations (370, 600, and 1000 ppmv), temperature (6°C, 12°C, and 18°C) and invertebrate biodiversity on MPB biomass in experimental systems. We assembled communities of three common grazing invertebrates (Hydrobia ulvae, Corophium volutator and Hediste diversicolor) in monoculture and in all possible multispecies combinations. This experimental design specifically addresses interactions between the selected climate change variables and any ecological consequences caused by changes in species composition or richness.     

Effect of Early Postpartum Ovulation on Fertility in Dairy Cows

Our objectives were to determine the effects of early ovulation on fertility and uterine health of dairy cows. Four hundred and forty‐five Holstein cows (185 primiparous and 260 multiparous) from five herds were used. Blood samples were collected at 21, 35 and 49 days in milk (DIM) and cows were considered to be cyclic at 21 DIM (Cyc21) if serum progesterone (P4) concentration was above 1 ng/ml, cyclic by 49 DIM (Cyc49) if P4 concentration was above 1 ng/ml at 35 or 49 DIM, or not cyclic (NotCyc) if P4 concentration was below 1 ng/ml at all sample times. Endometrial cytology for diagnosis of subclinical endometritis was examined at 49 DIM in a subset of 414 cows. Cows in the group Cyc21 had increased hazard of insemination, for the first service, compared with cows in Cyc49 [hazard ratio (HR) = 1.40; 95% CI = 1.10–1.79; p = 0.006] and NotCyc (HR = 2.07; 95% CI = 1.52–2.82; p < 0.001). Cows in the Cyc49 group also had increased hazard of insemination compared with cows in the NotCyc group (HR = 1.47; 95% CI = 1.13–1.93; p = 0.005). Median days to insemination were, respectively, 71, 76 and 96 for cows in Cyc21, Cyc49 and NotCyc groups. Cows in Cyc21 had greater first service pregnancy per AI than Cyc49 [38.6 vs 28.1%; adjusted odds ratio (AOR) = 1.67; 95% CI = 1.01–2.75; p = 0.04] and NotCyc (38.6 vs 23.6%; AOR = 2.08; 95% CI = 1.08–4.00; p = 0.03). Pregnancy per AI was similar in Cyc49 and NotCyc cows (28.1 vs 23.6%; AOR = 1.25; 95% CI = 0.70–2.24; p = 0.45). Cows in Cyc21 had increased hazard of pregnancy up to 300 DIM compared with Cyc49 (HR = 1.52; 95% CI = 1.17–1.96; p = 0.002) and NotCyc (HR = 1.98; 95% CI = 1.41–2.78; p < 0.001). Cows in Cyc49 tended to have increased hazard of pregnancy compared with NotCyc (HR = 1.31; 95% CI = 0.96–1.77; p = 0.09). Median days to pregnancy were, respectively, 103, 147 and 173 for cows in Cyc21, Cyc49 and NotCyc groups. Cows in the Cyc21 group had decreased prevalence of subclinical endometritis compared with cows in the NotCyc group (29.9 vs 43.7%; AOR = 0.53; 95% CI = 0.29–0.97; p = 0.04); however, the prevalence did not differ from the Cyc49 group (29.9 vs 39.1%; AOR = 0.68; 95% CI = 0.41–1.14; p = 0.15). Cyc49 cows had similar prevalence of subclinical endometritis compared with NotCyc cows (AOR = 0.77; 95% CI = 0.46–1.29; p = 0.32). Early postpartum ovulation was associated with improved uterine health and fertility.     

Effects of Escherichia coli Shiga-like toxins (verotoxins) in pigs.

Escherichia coli K12 strains TB1(pCG5), with the genes for Shiga-like toxin IIv from an edema disease isolate of E. coli and TB1(pCG5-1), with the toxin genes inactivated by transposon mutagenesis, were used to test the hypothesis that Shiga-like toxin IIv was the same as edema disease principle. Ammonium sulfate precipitated culture supernatants from the pair of E. coli K12 strains and from a wild edema disease isolate of E. coli (E145) were tested for their ability to induce signs and lesions of edema disease in intravenously inoculated weaned pigs. Similar preparations from E. coli which produce Shiga-like toxins I and II were also tested. Preparations from E. coli TB1 (pCG5) and E145 contained high levels of Shiga-like toxin IIv and induced signs and lesions similar to those seen in edema disease, whereas preparations from E. coli TB1 (pCG5-1) failed to induce signs or lesions of edema disease. All Shiga-like toxin preparations produced delayed neurological signs, fibrinoid necrosis of arterioles and hemorrhages in the cerebellum of pigs. High doses of Shiga-like toxin IIv were associated with superficial necrosis of the colonic epithelium and vasculitis. Shiga-like toxins I and II resulted in kidney lesions but no enteric pathology. Shiga-like toxin II preparations had the lowest median lethal dose for pigs, Shiga-like toxin IIv was intermediate and Shiga-like toxin I was the least toxic.